Physical Activity Modifies the Associations between Genetic Variants and Blood Pressure in European Adolescents Augusto Cesar Ferreira de Moraes, PhD1,2,3, Juan Miguel Fernandez-Alvira, PhD2,3, Heraclito Barbosa Carvalho, PhD1, Aline Meirhaeghe, PhD4, Jean Dallongeville, PhD4, Anthony Kafatos, PhD5, Ascension Marcos, PhD6, Denes Molnar, PhD7, Yannis Manios, PhD8, Jonatan R. Ruiz, PhD9, Idoia Labayen, PhD10, Kurt Widhalm, PhD11, Christina Breidenassel, PhD12, Marcela Gonzalez-Gross, PhD12,13, and Luis A. Moreno, PhD2,3,* We hypothesized that physical activity and sedentary behavior could modify the associations between known genetic variants blood pressure-associated genes in European adolescents. Meeting current physical activity recommendations ($60 minutes/day) was able attenuate the deleterious effect of the NOS3 rs3918227 polymorphism on systolic blood pressure in European adolescents. (J Pediatr 2014;-:---).
H
ypertension is a major public health problem and risk factor for cardiovascular disease.1 Recent studies have suggested an association between blood pressure (BP) levels and certain genetic loci, with special potential gene regulatory mechanisms at the MTHFR and NOS3 loci.2,3 In addition, BP is known to be affected by several environmental factors, among which are physical activity and sedentary behaviors, as predictors of higher BP.4 As genetic influence on BP would have been already observed during adolescence,5 determining to what extent BP-related genes are modulated by environmental factors such as physical activity and sedentary behaviors during adolescence could help to further understand and prevent high BP at early stages. Thus, we hypothesized that physical activity and sedentary behavior levels could modify the associations between genetic variants known BP-associated variants in European adolescents (Figure 1; available at www.jpeds.com).
Methods A total of 3528 adolescents (1845 girls) from 9 European countries participated in the Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) study. A detailed description of the HELENA methodology has been published elsewhere.6,7 The Human Research Review Committees of the centers involved approved the protocol following the Helsinki Declaration rules. A total of 1009 adolescents (12.5-17.5 years old; 532 girls), with valid data on BP levels, physical activity measured by accelerometry, sedentary behavior assessed by questionnaire, and a complete set of confounders were included in the analyses of this study.
BP DBP HELENA SBP
Blood pressure Diastolic blood pressure Healthy Lifestyle in Europe by Nutrition in Adolescence Systolic blood pressure
Systolic BP (SBP) and diastolic BP (DBP) were measured by an arm BP oscillometric monitor device (OMRON M6, HEM 70001; Omron, Kyoto, Japan) and following the recommendations for adolescent populations,8 complete data collection procedures have been described in earlier.9 Independent Variables Genotyping. Fasting blood samples were collected by venipuncture after a 10-hour overnight fast, as previously described.10 For the purpose of the present study, 6 single nucleotide polymorphisms in 3 genes previously associated with hypertension3,11 were selected (rs1205 and rs1130864 in CRP, rs1801131 and rs1537516 in MTHFR, and rs1800779 and rsc3918227 in NOS3). Genotyping success rates were greater than 99.7%. The genotype distributions,
From the 1Department of Preventive Medicine, School of Medicine of the University ~o Paulo, Sa ~o Paulo, SP, Brazil; 2GENUD (Growth, Exercise, Nutrition, and of Sa Development) Research Group, Universidad de Zaragoza, Zaragoza, Spain; 3Faculty of Health Sciences of the University of Zaragoza, Zaragoza, Spain; 4INSERM, U744, Institut Pasteur de Lille, Univ Lille Nord de France; UDSL, Lille, France; 5Preventive Medicine and Nutrition Unit, University of Crete School of Medicine, Heraklion, Crete, Greece; 6Immunonutrition Research Group, Department of Metabolism and Nutrition, Food Science and Technology and Nutrition Institute, Spanish National Research Council, Madrid, Spain; 7Department of Pediatrics, Medical Faculty, cs, Pe cs, Hungary; 8Department of Nutrition and Dietetics, Harokopio University of Pe University, Athens, Greece; 9Promoting Fitness and Health through Physical Activity Research Group, Department of Physical Education and Sports, Faculty of Sport Sciences, University of Granada, Granada, Spain; 10Department of Nutrition and Food Science, University of the Basque Country, Vitoria, Spain; 11Department of €r Pediatrics, Private Medical University Salzburg, Salzburg, Austria; 12Institut fu €hrungs-und Lebensmittelwissenschaften–Humanerna €hrung, Rheinische Erna 13 €t Bonn, Bonn, Germany; and Department of Health Friedrich-Wilhelms-Universita and Human Performance, Faculty of Physical Activity and Sport-INEF, Technical University of Madrid, Madrid, Spain *L.M. is a visiting professor at the Department of Preventive Medicine, School of ~o Paulo, Sa ~o Paulo, SP, Brazil. Medicine of the University of Sa The HELENA Study was supported by the European Community Sixth Research, Technological Development and Demonstration Framework Programme (FOODCT-2005-007034), the Spanish Ministry of Health: Maternal, Child Health, and Development Network (number RD08/0072), the Spanish Ministry of Education (EX€rt-Lungfonden (20090635). The 2008-0641 and RYC-2010-05957), and the Hja GENUD Research Group was funded in part by the European Regional Develop~o Paulo ment Fund (MICINN-FEDER). A.dM. received a scholarship from Sa Research Foundation (2011/11137-1 and 2011/20662-2). L.M. received a scholarship of visiting professor from Brazilian government by Science without Borders Program by National Counsel of Technological and Scientific Development and Coordination of Improvement of Higher Education Personnel (007/2012). The authors declare no conflicts of interest. 0022-3476/$ - see front matter. Copyright ª 2014 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.jpeds.2014.07.007
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Vol. -, No. -
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respected the Hardy-Weinberg equilibrium, for all polymorphisms (P > .25).
To test for the existence of an interaction between each SNP and lifestyle variables, the same models were applied by adding the SNPs lifestyle cross-product (eg, rs1205 physical activity) in the model. Significance was set at P values of 4 hours/day) than girls. Boys had also higher waist circumference and SBP than girls, whereas girls had higher DBP levels than boys (Table). The CRP rs1130864 and NOS3 rs3918227 SNPs were associated with both BP levels. Indeed, adolescents carrying the minor allele of rs1130864 had higher DBP (P = .003) and SBP (P = .041) levels than carriers of the major allele. Similarly, adolescents carrying the minor allele of rs3918227 had higher SBP (P = .026) and DBP (P = .026) levels than carriers of the major allele. Furthermore, the adolescents bearing the minor allele of the MTHFR rs1537516 SNP had higher values of DBP than the other adolescents (P = .017; Figure 2 available at www.jpeds.com). We did not detect any significant interaction between the levels of sedentary behavior, polymorphisms, and BP levels
Statistical Analyses Descriptive analyses are presented as means and percentages, 95% CIs. The impact of the 6 polymorphisms (using the additive model) on DBP and SBP levels were estimated by multilevel linear regression models using mixed effects intercept adjusted for potential confounders. City was used as context variable. Table. Characteristics of the subjects in the HELENA study Variables Age (y) Education mother Lower education Lower secondary education Higher secondary education University degree Tanner Stage 1 and 2 (prepubertal) 3 and 4 (pubertale) 5 (postpubertal) MVPA
H
ypertension is a major public health problem and risk factor for cardiovascular disease.1 Recent studies have suggested an association between blood pressure (BP) levels and certain genetic loci, with special potential gene regulatory mechanisms at the MTHFR and NOS3 loci.2,3 In addition, BP is known to be affected by several environmental factors, among which are physical activity and sedentary behaviors, as predictors of higher BP.4 As genetic influence on BP would have been already observed during adolescence,5 determining to what extent BP-related genes are modulated by environmental factors such as physical activity and sedentary behaviors during adolescence could help to further understand and prevent high BP at early stages. Thus, we hypothesized that physical activity and sedentary behavior levels could modify the associations between genetic variants known BP-associated variants in European adolescents (Figure 1; available at www.jpeds.com).
Methods A total of 3528 adolescents (1845 girls) from 9 European countries participated in the Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) study. A detailed description of the HELENA methodology has been published elsewhere.6,7 The Human Research Review Committees of the centers involved approved the protocol following the Helsinki Declaration rules. A total of 1009 adolescents (12.5-17.5 years old; 532 girls), with valid data on BP levels, physical activity measured by accelerometry, sedentary behavior assessed by questionnaire, and a complete set of confounders were included in the analyses of this study.
BP DBP HELENA SBP
Blood pressure Diastolic blood pressure Healthy Lifestyle in Europe by Nutrition in Adolescence Systolic blood pressure
Systolic BP (SBP) and diastolic BP (DBP) were measured by an arm BP oscillometric monitor device (OMRON M6, HEM 70001; Omron, Kyoto, Japan) and following the recommendations for adolescent populations,8 complete data collection procedures have been described in earlier.9 Independent Variables Genotyping. Fasting blood samples were collected by venipuncture after a 10-hour overnight fast, as previously described.10 For the purpose of the present study, 6 single nucleotide polymorphisms in 3 genes previously associated with hypertension3,11 were selected (rs1205 and rs1130864 in CRP, rs1801131 and rs1537516 in MTHFR, and rs1800779 and rsc3918227 in NOS3). Genotyping success rates were greater than 99.7%. The genotype distributions,
From the 1Department of Preventive Medicine, School of Medicine of the University ~o Paulo, Sa ~o Paulo, SP, Brazil; 2GENUD (Growth, Exercise, Nutrition, and of Sa Development) Research Group, Universidad de Zaragoza, Zaragoza, Spain; 3Faculty of Health Sciences of the University of Zaragoza, Zaragoza, Spain; 4INSERM, U744, Institut Pasteur de Lille, Univ Lille Nord de France; UDSL, Lille, France; 5Preventive Medicine and Nutrition Unit, University of Crete School of Medicine, Heraklion, Crete, Greece; 6Immunonutrition Research Group, Department of Metabolism and Nutrition, Food Science and Technology and Nutrition Institute, Spanish National Research Council, Madrid, Spain; 7Department of Pediatrics, Medical Faculty, cs, Pe cs, Hungary; 8Department of Nutrition and Dietetics, Harokopio University of Pe University, Athens, Greece; 9Promoting Fitness and Health through Physical Activity Research Group, Department of Physical Education and Sports, Faculty of Sport Sciences, University of Granada, Granada, Spain; 10Department of Nutrition and Food Science, University of the Basque Country, Vitoria, Spain; 11Department of €r Pediatrics, Private Medical University Salzburg, Salzburg, Austria; 12Institut fu €hrungs-und Lebensmittelwissenschaften–Humanerna €hrung, Rheinische Erna 13 €t Bonn, Bonn, Germany; and Department of Health Friedrich-Wilhelms-Universita and Human Performance, Faculty of Physical Activity and Sport-INEF, Technical University of Madrid, Madrid, Spain *L.M. is a visiting professor at the Department of Preventive Medicine, School of ~o Paulo, Sa ~o Paulo, SP, Brazil. Medicine of the University of Sa The HELENA Study was supported by the European Community Sixth Research, Technological Development and Demonstration Framework Programme (FOODCT-2005-007034), the Spanish Ministry of Health: Maternal, Child Health, and Development Network (number RD08/0072), the Spanish Ministry of Education (EX€rt-Lungfonden (20090635). The 2008-0641 and RYC-2010-05957), and the Hja GENUD Research Group was funded in part by the European Regional Develop~o Paulo ment Fund (MICINN-FEDER). A.dM. received a scholarship from Sa Research Foundation (2011/11137-1 and 2011/20662-2). L.M. received a scholarship of visiting professor from Brazilian government by Science without Borders Program by National Counsel of Technological and Scientific Development and Coordination of Improvement of Higher Education Personnel (007/2012). The authors declare no conflicts of interest. 0022-3476/$ - see front matter. Copyright ª 2014 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.jpeds.2014.07.007
1
THE JOURNAL OF PEDIATRICS
Vol. -, No. -
www.jpeds.com
respected the Hardy-Weinberg equilibrium, for all polymorphisms (P > .25).
To test for the existence of an interaction between each SNP and lifestyle variables, the same models were applied by adding the SNPs lifestyle cross-product (eg, rs1205 physical activity) in the model. Significance was set at P values of 4 hours/day) than girls. Boys had also higher waist circumference and SBP than girls, whereas girls had higher DBP levels than boys (Table). The CRP rs1130864 and NOS3 rs3918227 SNPs were associated with both BP levels. Indeed, adolescents carrying the minor allele of rs1130864 had higher DBP (P = .003) and SBP (P = .041) levels than carriers of the major allele. Similarly, adolescents carrying the minor allele of rs3918227 had higher SBP (P = .026) and DBP (P = .026) levels than carriers of the major allele. Furthermore, the adolescents bearing the minor allele of the MTHFR rs1537516 SNP had higher values of DBP than the other adolescents (P = .017; Figure 2 available at www.jpeds.com). We did not detect any significant interaction between the levels of sedentary behavior, polymorphisms, and BP levels
Statistical Analyses Descriptive analyses are presented as means and percentages, 95% CIs. The impact of the 6 polymorphisms (using the additive model) on DBP and SBP levels were estimated by multilevel linear regression models using mixed effects intercept adjusted for potential confounders. City was used as context variable. Table. Characteristics of the subjects in the HELENA study Variables Age (y) Education mother Lower education Lower secondary education Higher secondary education University degree Tanner Stage 1 and 2 (prepubertal) 3 and 4 (pubertale) 5 (postpubertal) MVPA
